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KMID : 0613820000100040368
Journal of Life Science
2000 Volume.10 No. 4 p.368 ~ p.373
An Effective Method for the Concentration and Detection of Enteroviruses from Water Samples by Combined Cell Culture-Polymerase Chain Reaction

Abstract
Enteroviruses in the environment pose a public health risk because they can be transmitted via the fecal-oral route through contaminated water, and low numbers are able to initiate an infection in humans. Because the levels of viruses typically found in environmental water and drinking water are low, they must be concentrated from hundreds to thousands of liters of water. Therefore, the main goal of this study was the development of a rapid, simple and efficient procedure to concentrate, isolate and detect enteroviruses from environmental water samples. Viruses were first concentrated by adsorption to 1 MDS cartridge filter and then eluted with approximately 0.5 liter of 1.5% beef extract/0.05M glycin(pH 9.4). In this study, several procedures to concentrate and purify intact viruses from beef extract obtained from the adsorbent filters were tested. Among them, organic floccuration was the best reliable method for reconcentration. sample volume could be reduced to 200¡­400 folds and the efficiency of virus recovery through the procedure was over 72%. Finally, the samples were filtered through a membrane disk filter and then analyzed by either the plaque assay or combined cell culture-polymerase chain reaction.
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